17β-estradiol attenuates ovariectomy‑induced bone deterioration through the suppression of the ephA2/ephrinA2 signaling pathway

Liu,Lianyong; Zhou,Lin; Yang,Xiaorong; Liu,Qi; Yang,Ling; Zheng,Chao; Zhao,Yongling; Zhang,Zhenlin; Luo,Xiaohong

doi:10.3892/mmr.2017.8042

17β-estradiol attenuates ovariectomy‑induced bone deterioration through the suppression of the ephA2/ephrinA2 signaling pathway

Authors:
- Lianyong Liu
- Lin Zhou
- Xiaorong Yang
- Qi Liu
- Ling Yang
- Chao Zheng
- Yongling Zhao
- Zhenlin Zhang
- Xiaohong Luo
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Affiliations: Medical College of Soochow University, Suzhou, Jiangsu 215123, P.R. China, Department of Endocrinology, Punan Hospital of Pudong New District, Shanghai 200125, P.R. China, Metabolic Bone Disease and Genetic Research Unit, Department of Osteoporosis and Bone Diseases, Shanghai Key Clinical Center for Metabolic Disease, Sixth People's Hospital Affiliated to Shanghai Jiao Tong University, Shanghai 200233, P.R. China
Published online on: November 14, 2017 https://doi.org/10.3892/mmr.2017.8042
Pages: 1609-1616
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to investigate whether 17β‑estradiol (E2) exerts protective effects on bone deterioration induced by ovariectomy (OVX) through the ephA2/ephrinA2 signaling pathway in rats. Female rats were subjected to OVX, sham surgeryor OVX+E2 treatment. Levels of biomarkers were measured in serum and urine. Hematoxylin and eosin staining was performed on paraffin‑embedded bone sections. Expression of genes and proteins was analyzed by reverse transcription‑quantitative polymerase chain reaction and western blotting, respectively. Bone mineral density (BMD) was analyzed by dual‑energy X‑ray absorptiometry. Trabecular bone microarchitecture was also evaluated. Osteoclastogenesis was induced by in vitro culturing with mouse receptor activator of nuclear factor κB ligand (RANKL) and macrophage colony‑stimulating factor 1. small interfering RNA was designed to knockdown ehpA2 receptor and its ligand ephrinA2. Results of the present study demonstrated that E2 had suppressive effects on OVX‑induced body weight gain and bone turnover factors in serum and urine. E2 inhibited the bone resorption function of osteoclasts by inhibiting the production of tartrate‑resistant acid phosphatase‑5b and RANKL, and induced bone formation function of osteoblasts by prompting runt‑related transcription factor 2, Sp7 transcription factor and collagen alpha‑1(I) chain expression in bone marrow cells. E2 treatment significantly increased the tibia BMD and prevented the deterioration of trabecular microarchitecture compared with the OVX group. Moreover, E2 significantly decreased the OVX‑stimulated expression of ephA2 and ephrinA2. EphA2 or ephrin A2 knockdown significantly suppressed osteoclastogenesis in vitro. In conclusion, E2 can attenuate OVX‑induced bone deterioration partially through the suppression of the ephA2/ephrinA2 signaling pathway. Therefore EphA2/ephrinA2 signaling pathway may be a potential target for osteoporosis treatment.

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1	Jin S, Yan Z, Yang T, Liu S, Liang W and Hui Y: Eph-ephrin bidirectional signalling: A promising approach for osteoporosis treatment. J Med Hypotheses Ideas. 7:40–42. 2013. View Article : Google Scholar
2	Karsenty G and Wagner EF: Reaching a genetic and molecular understanding of skeletal development. Dev Cell. 2:389–406. 2002. View Article : Google Scholar
3	Matsuo K and Irie N: Osteoclast-osteoblast communication. Arch Biochem Biophys. 473:201–209. 2008. View Article : Google Scholar
4	Rachner TD, Khosla S and Hofbauer LC: Osteoporosis: Now and the future. Lancet. 377:1276–1287. 2011. View Article : Google Scholar :
5	Sarrel PM, Lufkin EG, Oursler MJ and Keefe D: Estrogen actions in arteries, bone and brain. Sci Med. 1:441994.
6	Nappi C, Bifulco G, Tommaselli GA, Gargano V and Di Carlo C: Hormonal contraception and bone metabolism: A systematic review. Contraception. 86:606–621. 2012. View Article : Google Scholar
7	Riggs BL: The mechanisms of estrogen regulation of bone resorption. J Clin Invest. 106:1203–1204. 2000. View Article : Google Scholar :
8	Eastell R: Role of oestrogen in the regulation of bone turnover at the menarche. J Endocrinol. 185:223–234. 2005. View Article : Google Scholar
9	Zhou S, Turgeman G, Harris SE, Leitman DC, Komm BS, Bodine PV and Gazit D: Estrogens activate bone morphogenetic protein-2 gene transcription in mouse mesenchymal stem cells. Mol Endocrinol. 17:56–66. 2003. View Article : Google Scholar
10	Hong L, Colpan A and Peptan IA: Modulations of 17-beta estradiol on osteogenic and adipogenic differentiations of human mesenchymal stem cells. Tissue Eng. 12:2747–2753. 2006. View Article : Google Scholar
11	Hong L, Colpan A, Peptan IA, Daw J, George A and Evans CA: 17-Beta estradiol enhances osteogenic and adipogenic differentiation of human adipose-derived stromal cells. Tissue Eng. 13:1197–1203. 2007. View Article : Google Scholar
12	Irie N, Takada Y, Watanabe Y, Matsuzaki Y, Naruse C, Asano M, Iwakura Y, Suda T and Matsuo K: Bidirectional signaling through ephrinA2-EphA2 enhances osteoclastogenesis and suppresses osteoblastogenesis. J Biol Chem. 284:14637–14644. 2009. View Article : Google Scholar :
13	Difford J: A simplified method for the preparation of methyl methacrylate embedding medium for undecalcified bone. Med Lab Technol. 31:79–81. 1974.
14	Baron R, Vignery A, Neff L, Silverglate A and Maria Santa A: Processing of undecalcified bone specimens for bone histomorphometryBone Histomorphometry: Teachniques and Interpretation. Recker R: CRC Press; Boca Raton, FL: 1983
15	Witwicka H, Hwang SY, Reyes-Gutierrez P, Jia H, Odgren PE, Donahue LR, Birnbaum MJ and Odgren PR: Studies of OC-STAMP in osteoclast fusion: A new knockout mouse model, rescue of cell fusion and transmembrane topology. PLoS One. 10:e01282752015. View Article : Google Scholar :
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
17	Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, et al: Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 89:755–764. 1997. View Article : Google Scholar
18	Ortuño MJ, Susperregui AR, Artigas N, Rosa JL and Ventura F: Osterix induces Col1a1 gene expression through binding to Sp1 sites in the bone enhancer and proximal promoter regions. Bone. 52:548–556. 2013. View Article : Google Scholar
19	Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Lüthy R, Nguyen HQ, Wooden S, Bennett L, Boone T, et al: Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell. 89:309–319. 1997. View Article : Google Scholar
20	Kong YY, Yoshida H, Sarosi I, Tan HL, Timms E, Capparelli C, Morony S, Oliveira-dos-Santos AJ, Van G, Itie A, et al: OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature. 397:315–323. 1999. View Article : Google Scholar
21	Shuid AN, Ping LL, Muhammad N, Mohamed N and Soelaiman IN: The effects of Labisia pumila var. alata on bone markers and bone calcium in a rat model of post-menopausal osteoporosis. J Ethnopharmacol. 133:538–542. 2011. View Article : Google Scholar
22	Zhao X, Wu ZX, Zhang Y, Yan YB, He Q, Cao PC and Lei W: Anti-osteoporosis activity of cibotium barometz extract on ovariectomy-induced bone loss in rats. J Ethnopharmacol. 137:1083–1088. 2011. View Article : Google Scholar
23	Notomi T, Okimoto N, Okazaki Y, Nakamura T and Suzuki M: Tower climbing exercise started 3 months after ovariectomy recovers bone strength of the femur and lumbar vertebrae in aged osteopenic rats. J Bone Miner Res. 18:140–149. 2003. View Article : Google Scholar
24	Davidge ST, Zhang Y and Stewart KG: A comparison of ovariectomy models for estrogen studies. Am J Physiol Regul Integr Comp Physiol. 280:R904–R907. 2001.
25	Redick JH, Nussbaum AI and Mook DG: Estradiol induced suppression of feeding in the female rat: Dependence on body weight. Physiol Behav. 10:543–547. 1973. View Article : Google Scholar
26	Bahlous A, Kalai E, Salah Hadj M, Bouzid K and Zerelli L: Biochemical markers of bone remodeling: Recent data of their applications in managing postmenopausal osteoporosis. Tunis Med. 84:751–757. 2006.(In French).
27	Zheng MH, Lau TT, Prince R, Criddle A, Wysocki S, Beilharz M, Papadimitriou JM and Wood DJ: 17 beta-estradiol suppresses gene expression of tartrate-resistant acid phosphatase and carbonic anhydrase II in ovariectomized rats. Calcif Tissue Int. 56:166–169. 1995. View Article : Google Scholar
28	Park JA, Ha SK, Kang TH, Oh MS, Cho MH, Lee SY, Park JH and Kim SY: Protective effect of apigenin on ovariectomy-induced bone loss in rats. Life Sci. 82:1217–1223. 2008. View Article : Google Scholar
29	Shimizu E, Tamasi J and Partridge NC: Alendronate affects osteoblast functions by crosstalk through EphrinB1-EphB. J Dent Res. 91:268–274. 2012. View Article : Google Scholar :
30	Matsuo K: Eph and ephrin interactions in bone. Adv Exp Med Biol. 658:95–103. 2010. View Article : Google Scholar
31	Matsuo K and Otaki N: Bone cell interactions through Eph/ephrin: Bone modeling, remodeling and associated diseases. Cell Adh Migr. 6:148–156. 2012. View Article : Google Scholar :